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학술발표회초록보기

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  • 09월 03일 23시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제122회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Single-molecule FRET study on CRISPR-Cas12a endonuclease

등록일
2018년 9월 3일 18시 27분 23초
접수번호
2208
발표코드
LIFE2-3 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
금 09시 : 50분
발표형식
심포지엄
발표분야
Life Chemistry - Recent Trends in Genome Editing Technique
저자 및
공동저자
Sanghwa Lee
Advanced Photonics Research Institute, Gwangju Institute of Science and Technology, Korea

Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to type II CRISPR–Cas9 for genome editing. Previous studies have revealed that Cas12a has unique features distinct from Cas9, but the detailed mechanisms of DNA target recognition and cleavage by Cas12a are still unclear. In this work, we directly observe this entire process by using single-molecule fluorescence assays to study Cas12a from Acidaminococcus sp. (AsCas12a). our results reveal that a stable R-loop formation, which is the main determinant of target recognition, is efficiently established only when the seed matches up to 17 base pairs, and AsCas12a ribonucleoproteins induce cleavage in the two DNA strands in a well-defined order, beginning with the non-target strand. Furthermore, the protospacer-adjacent motif (PAM) for AsCas12a makes only a limited contribution of DNA unwinding during R-loop formation and shows a negligible role in the process of DNA cleavage, in contrast to the Cas9 PAM.


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